Useful in Sandwich ELISA for Quantitative Detection of Antigen. Aliquot 0.1ml per well of the 2000pg/ml,1000pg/ml, 500pg/ml, 250pg/ml, 125pg/ml, 62.5pg/ml, 31.2pg/ml human Nesfatin-1 standard solutions into the precoated 96-well plate. Add 0.1ml of the sample diluent buffer into the control well (Zero well). Add 0.1ml of each properly diluted sample of human cell culture supernates or serum to each empty well. It is recommended that each human Nesfatin-1 standard solution and each sample be measured in duplicate. Seal the plate with the cover and incubate at 37°C for 90 min.